Biocidal methods employing perhalocoumalin derivatives

ABSTRACT

A COMPOUND SELECTED FROM THE GROUP CONSISTING OF COMPOUNDS OF THE FORMULAE   2-(O=),3,4,5-TRI(X-),6-((R)N-Z-)-2H-PYRAN, AND   (R)N-Z-CO-CH(-Z(-R)N)-C(-X)=C(-X)-CO-Z(-R)N   WHEREIN X IS A HALOGEN CHOSEN FROM THE GROUP CONSISTING OF FLUORINE, CHLORINE, BROMINE, AND IODINE, AND MIXTURES THEREOF. (R)NZ IS A SUBSTITUENTS IN WHICH THE FOLLOWING RELATINGSHIP EXISTS:   IF N IS- Z MAY BE0 BROMINE, IODINE, FLUORINE. 1 OXYGEN, SULFUR, SELENIUM. 2 NITROGEN, ARSENIC. 3 CARBON.   AND WHEREIN R IS SELECTED FROM THE GROUP CONSISTING OF HYDROGEN; ALKYL GROUPS OF 1 TO 18 CARBON ATOMS IN ANY OF THEIR ISOMERIC FORMS, PROVIDED THAT WHEN Z IS OXYGEN THE ALKYL GROUPS FROM WHICH R MAY BE SELECTED CONTAIN FROM 6 TO 18 CARBON ATOMS; SUBSTITUTED ALKYL GROUPS OF 1 TO 18 CARBON ATOMS WHEREIN THE SUBSTITUENTS ARE SELECTED FROM THE GROUP CONSISTING OF FLUORINE, CHLORINE, BROMINE, IODINE, SULFUR, OXYGEN, PHOSPHOURS, NITROGEN, AND SILICON; ALICYCLIC GROUPS CONTAINING FROM 5 TO 18 CARBON ATOMS; HETEROCYCLIC GROUPS CONTAINING FROM 5 TO 18 CARBON ATOMS WHEREIN THE HETERO ATOM(S) IS SELECTED FROM THE GROUP CONSISTING OF OXYGEN, SULFUR, NITROGEN, PHOSPHORUS, AND SILICON; AND AROMATIC AND ARALKYL GROUPS CONTAINING FROM 6 TO 18 CARBON ATOMS. THE COMPOUNDS ARE USEFUL AS PESTICIDES, BACTERICIDES, FUNGICIDES, NEMATOCIDES, INSECTICIDES AND MITICIDES.

United States Patent 3,755,591 Patented Aug. 28, 1973 US. Cl. 424-279 16 Claims ABSTRACT OF THE DISCLOSURE A compound selected from the group consisting of compounds of the formulae wherein X is a halogen chosen from the group consisting of fluorine, chlorine, bromine, and iodine, and mixtures thereof. (R) Z is a substituents in which the following relationship exists:

If n is Z may be 0 Bromine, iodine, fluorine. 1 Oxygen, sulfur, selenium. 2 Nitrogen, arsenic. 3 Carbon.

and wherein R is selected from the group consisting of hydrogen; alkyl groups of l to 18 carbon atoms in any of their isomeric forms, provided that when Z is oxygen the alkyl groups from which R may be selected contain from 6 to 18 carbon atoms; substituted alkyl groups of 1 to 18 carbon atoms wherein the substituents are selected from the group consisting of fluorine, chlorine, bromine, iodine, sulfur, oxygen, phosphorus, nitrogen, and silicon; alicyclic groups containing from 5 to 18 carbon atoms; heterocyclic groups containing from 5 to 18 carbon atoms wherein the hetero atom(s) is selected from the group consisting of oxygen, sulfur, nitrogen, phosphorus, and silicon; and aromatic and aralkyl groups containing from 6 to 18 carbon atoms.

The compounds are useful as pesticides, bactericides, fungicides, nematocides, insecticides and miticides.

This is a division of our parent application, Ser. No. 586,056, filed Oct. 12, 1966, now US. Pat. 3,528,995 issued Sept. 15, 1970.

This invention relates to new compounds, to processes for their preparation, and to the uses to which said compounds may be put. More specifically the invention relates to novel substituted halogenated coumalins and the derivatives thereof, to novel methods for producing said compounds, and to the fungicidal, bactericidal, insecticidal, pesticidal uses of said compounds.

In accordance with this invention there are provided compounds of the formulae:

wherein X is a halogen chosen from the group consisting of fluorine, chlorine, bromine, and iodine and mixtures thereof (R),,Z is a substituent in which the following relationship exists:

If n is Z may \be- 0 Bromine, iodine, fluorine. l Oxygen, sulfur, selenium. 2 Nitrogen, arsenic. 3 Carbon.

and wherein R is selected from the group consisting of hydrogen; alkyl groups of 1 to 18 carbon atoms in any of their isomeric forms provided that when Z is oxygen the alkyl groups from which R may be selected contain from 6 to 18 carbon atoms; substituted alkyl groups of 1 to 18 carbon atoms wherein the substituents are selected from the group consisting of fluorine, chlorine, bromine, iodine, sulfur, oxygen, phosphorus, nitrogen, and silicon; alicyclic groups containing from 5 to 18 carbon atoms; heterocyclic groups containing from 5 to 18 carbon atoms wherein the hetero atom(s) is selected from the group consisting of oxygen, sulfur, nitrogen, phosphorus, and silicon; and aromatic and aralkyl groups containing from 6 to 18 carbon atoms.

In a preferred embodiment of the invention, Z is sulfur, X is chlorine, and R is an alkyl group of 1 to 12 carbon atoms, preferably of 1 to 5 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, and tert-butyl, most preferably of 1 carbon atom, methyl.

There is further provided a process for producing the aforementioned compounds by reacting a perhalocoumalin with a nucleophile in the presence of a base, as shown by the equations below:

wherein R, n, and X are as hereinbefore defined, provided that when n is 0, X is selected from the group consisting of fluorine, chlorine, and bromine, and the following relationship exists:

If X is Z may be Fluorine Chlorine, bromine, iodine. Chlorine Bromine, iodine. Bromine iodine.

The compounds so prepared exhibit a wide spectrum of pesticidal activity.

Any base which itself is not a strong nucleophilic precursor may be used as a coreagent. Thus tertiary amines may be used, and are the preferred class of bases, but a secondary or primary amine can be used if it is a relatively weak nucleophile precursor in comparison with the primary reacting species. Similarly, alkali or alkaline earth hydroxides, basic alkali salts, alkoxides, and salts of strong bases and weak acids may be used, provided their nucleophilicity is low compared to that of the reactant species.

A secondary amine which is a relatively strong nucleophile precursor may be used as the reactant species, and the reaction will proceed inasmuch as secondary amines act as both nucleophile precursors and bases, as illustrated by the equation shown below:

The perhalocoumalin is dissolved in a solvent, usually under ambient conditions, before it is made to react with the nucleophile precursor. In general, any polar or nonpolar solvent may be used. However, it is not desirable to use a solvent which itself will form a strong nucleophile with the base (such as, e.g., alcohols, amines) to avoid possible competing reactions.

The preferred solvents are ethers, e.g. tetrahydrofuran, diethylether, and the like. Other suitable solvents include benzene, acetone, methylene chloride, carbon disulfide, dimethylsulfoxide, and the like.

Once the perhalocoumalin is dissolved, it is made to react with the nucleophile precursor and base. When one is preparing the mono-substituted perhalocoumalin, about 1 mole of perhalocoumalin is reacted with each mole of nucleophile precursor and with an excess of base though it is preferred to use a substantially equirnolar amount of perhalocoumalin and nucleophile precursor. When one is preparing the tri-substituted derivative of glutaconic acid, about 1 mole of perhalocoumalin is reacted with 3 or more moles of nucleophile precursor and 2 or more moles of base, though it is preferred to use about 3 moles of nucleophile precursor for every mole of perhalocoumalin.

During the addition the temperature of the reaction mixture is maintained at to 180 degrees centrigrade, usually by cooling. A more preferred range is from 0-30 degrees centigrade, and an even more preferred range is from -30 degrees centigrade.

Pressure other than atmospheric is not required for this reaction. However, higher than atmospheric pressure may be used, when the nucleophile precursor is a gas at the reaction temperature.

Generally, the nucleophile precursor is added to the perhalocoumalin, followed by the addition of the base. However, where convenient, one may add the perhalocoumalin to the nucleophile precursor, and then add the base; alternatively, one may first add the base to the perhalocoumalin, and then add the nucleophile precursor thereto, and this method is preferred e.g., when the nucleophile precursor is a gas. When working with secondary amines, which act as both nucleophile precursors and bases, one preferably proceeds by adding them to the perhalocoumalins.

The reaction takes from about 10 minutes to 24 hours, being dependent upon the reactivity of the nucleophile generated from the precursor, the concentration of the nucleophile precursor, and the concentration of the perhalocoumalin. Often, the reaction rate is fast and it is completed upon the mixing of the reagents. After the addition is completed the reaction slurry is filtered and the solvent evaporated from the filtrate or the solvent can be evaporated, first and the resulting mixture triturated with water and the solids filtered off and dried. If the coumalin derivative is a liquid it can be isolated by filtration of the reaction slurr followed by evaporation of the solvent and distillation. Solid products may be purified by recrystallization from a solvent such as hexane, benzene, ethyl acetate, chloroform and the like.

In addition to making the tri-substituted derivatives of glutaconic acid in the manner described above, in, which all of the (R) Z groups are identical, one may alternatively make the mono-substituted perhalocoumalin using one nucleophile precursor, and then subject the monosubstituted perhalocoumalin using one nucleophile precursor, and then subject the mono-substituted perhalocoumalin to reaction with another nucleophile precursor. The equations presented below illustrate this.

0 0 base 01 I 0113311 base-H01 Cl =0 01 =0 C1 CI Cl 0 base 2n-O4H OH base H01 Cl =0 (1') C1 01 (H) CH3SC JOH(E=JZGOC4H,

Once the substituted perhalocoumalin products are made, various additional useful derivatives can be obtained. For example, -methylthio-3,4,5trichloro-2H- pyran-Z-one may be oxidized to the sulfone or sulfoxide, as illustrated below:

CH3 sons cs0 C c3 012 o 2011300311 2CH3CO2H or 0 c1 =0 The compounds of this invention are useful as chemical intermediates. They are also useful as fungicides, bactericides, and pesticides, as illustrated in Examples 13-33.

The following examples are presented as illustrative of the invention, and are not deemed to be limitative thereof. Unless otherwise specified, parts are by weight and temperatures are in degrees centigrade.

EXAMPLE 1 To a solution of 23.4 parts of perchlorocournalin and 10.1 parts of triethylamine (in parts of diethylether) were gradually added 5.0 parts of methyl mercaptan. The addition took place over a period of about 30 minutes and during said addition the temperature was maintained at 20-30 degrees centigrade. After completion of the addition, the solvent was evaporated and the residue was triturated with water, which water dissolved the amine hydrochloride and left behind the substituted coumalin. Said substituted coumalin was then recrystallized from hexane, quantitatively yielding yellow crystals which melted at 8889.5 degrees centigrade. Elemental analysis of the product disclosed that it contained 29.38 percent carbon, 1.23 percent hydrogen, 43.5 percent chlorine, and 12.95 percent sulfur, indicating that the compound had the formula SCH:

Cl =0 h EXAMPLES 2-11 In Examples 2-11 the basic procedure specified in Example l was followed, but different reactants and/or different concentrations of reactants were used, as specified in Table l.

7 SPRAY TESTS (FUNGICIDES) Early blight Tomato plants were sprayed with 100 milliliters of the chemical to be tested, dried, inoculated with spores of Altorernaria so'lam', and incubated in a moist chamber at 70 degrees Fahrenheit for 24 hours. Control plants were not sprayed wtih the chemical. Percent control was de termined by the formula:

(number of spots which develop on leaves of untreated tomato plant-number of spots which develop of leaves of treated plant) number of spots which develop on leaves of untreated plant Mildew Ten day old tendergreen bean plants were allowed to taken natural infection of Erysiphe polygoni from older infected plants. After the infection began to show on the leaves, the plants were sprayed with the chemical to be tested, dried, and kept in a greenhouse. Control plants were subjected to the aforementioned natural infection and kept in a greenhouse. When the control plants developed the disease, the test plants were removed from the greenhouse and inspected for infection.

SOIL FUNGICIDE TESTS Pythium test Soil naturally infested with Pythium species was treated with the chemical solution to be tested and allowed to stand 3 days before planting. Plastic pots 3.5 inches X 3.0 inches were filled with the soil, and the chemical was applied to the soil therein by drenching. After 3 days, ten pea seeds, Perfection variety, were planted in each pot. Percent emergence was recorded.

Rhizoctonia test Sterilized soil was inoculated with Strain #85 of Rhizoctonia solani, placed in 3.5 inches x 3.5 inches plastic pots, drenched with the chemical to be tested, and allowed to stand 3 days. Thereafter 5 Tendergreen bean seeds were planted in each pot. The number of plants developing infection was recorded.

Pusarium test Sterilized soil was inoculated with Fusarium oxysporum lycopersici, and treated with the chemical to be tested. Three days thereafter, five small tomato seedlings were planted in the soil. Fifteen days after said planting, the number of plants showing infection was recorded.

Sclerotium test Sterilized soil was inoculated with .S'clerotium rolfsii, treated with the chemical to be tested, and incubated in moist chambers at 70 degrees Fahrenheit for three days. Perfection variety peas were then planted in said soil, and the percent emergence was recorded.

Nematode test sterilized soil was inoculated with galls formed by Meloz'dogyne incognita on tomato roots, treated with the chemical to be tested, and allowed to stand for two or three days until it was dry enough for mixing. In such soil cucumber seeds were then planted, and the roots thereof were examined to days after planting for infection.

SPRAY TESTS (INSECTICIDES) Aphid test Ten-day-old Nasturtium plants were infested with back bean aphids (Aphis fabae Scop.) so that the first two leaves had from 50-100 aphids, sprayed with the chemical to be tested, and caged in a piece of tubing which is coated with talcum powder to prevent the escape of the aphids. Mortality was recorded by counting the number of dead aphids.

Mite test Young pole lirna bean plants (Sieva variety) were intested with 50 to adult mites (Tetranychus telariuu L.) on both primary leaves, sprayed 24 hours thereafter with the chemical to be tested, and placed in the greenhouse for from 24 to 48 hours, after which mortality was recorded by counting the number of dead mites.

Mexican bean beetle and army worm tests Primary leaves of lima bean plants were excised, dipped into a solution of the chemical to be tested, and dried. Either 5 larvae (fourth instar) of the Mexican bean beetle (Epz'lachna varivestia Muls.) or five larvae (second instar) of the southern armyworrn (Proa'enia eridania Cram), depending upon the test to be performed, were placed thereon, and the leaves were so enclosed that the insects could not escape. Mortality was recorded by counting the number of dead larvae, and the percent leaf consumed by the larvae was estimated.

House fly test Newly hatched adult house flies (Musca domestica L.) were, while under the action of carbon dioxide, immersed in the chemical solution to be tested, shaken for a few seconds, and screened out into a paper cup containing filter paper which was dipped in sugar solution, said cups then being covered to prevent escape of the flies. Ten flies were used per treatment. Data was taken on the number of flies killed in two hours, and killed in 24 hours.

Bactericidal test The bacteria to be tested were grown on agar slants for 20 hours. To a mixture of the chemical to be tested and nutrient agar was added two drops of the bacterial suspension, and this mixture was shaken and then poured onto sterilized Petri plates. The plates were incubated for 24 hours, and the bacterial growth noted.

EXAMPLE 14 The perhalocoumalin derivative of the formula S CH CH OH EXAMPLE 15 The perhalocoumalin derivative of the formula exhibited complete control of the bacteria Pseudomonas phaseolicola (Gram at a concentration of 255 parts per million, and at a concentration of 1000 parts per million killed 100 percent of the housefiies within 24 hours.

EXAMPLE 16 The perhalocoumalin derivative of the formula SCHaCH I CIA CH3 .l u,

exhibited complete control of the bacteria Staphylococcus aureus (Gram and Pseudomonas phaseolicola (Gram at concentrations of 255 parts per million and 76 parts per million, respectively, and at a concentration of 1000 parts per million killed 100 percent of the houseflies within 24 hours.

EXAMPLE 17 The perhalocoumalin derivative of the formula exhibited complete control of Staphylococcus aureus (Gram and Pseudomonas phaseolicola (Gram at a concentration of 255 parts per million. At a concentration of 1000 parts per million, it killed 20 percent of the southern armyworm, and at the same concentration it killed 100 percent of the houseflies within 24 hours.

EXAMPLE 18 The perhalocoumalin derivative of the formula C S Cg EXAMPLE 19 The perhalocoumalin derivative of the formula S CH \CHQ C1 exhibited complete control of Staphylococcus aureus Xanthomonas phasecoli (Gram at concentrations of 255, 38, and 255 parts per million, respectively. At 1000 parts per million concentration it killed 100 percent of the mites, and at the same concentration killed 100 percent of the h'ouseflies within two hours.

EXAMPLE 20 The perhalocoumalin derivative of the formula S CH:

exhibited percent control of Pythium at a concentration of 64 pounds per acre. It killed 29 percent of the aphids and percent of the mites at concentrations of 1000 and 290 parts per million, respectively. At a concentration of 1000 parts per million it killed percent of the houseflies within two hours.

EXAMPLE 21 The perhalocoumalin derivative of the formula killed 30 percent of the aphids and 70 percent of the mites at a concentration of 1000 parts per million. At the same concentration it killed 100 percent of the flies within two hours.

EXAMPLE 22 The perhalocoumalin derivative of the formula exhibited complete control of Pythium and Sclerotium at concentrations of 64 and 16 pounds per acre, respectively. It killed 38 percent of the aphids at a concentration of 1000 parts per million.

EXAMPLE 23 The derived coumalin product of the formula SCzHrSH killed 40 percent of the Southern armyworm and 100 percent of the Mexican bean beetle at concentrations of 1000 and 500 parts per million, respectively.

EXAMPLE 24 The derived coumalin product of the formula killed 60 percent of the Mexican bean beetle at a concentration of 1000 parts per million.

EXAMPLE 25 The derived coumalin product of the formula 0 CH S G CHC=Ci )SOH3 SCH: C1 C1 killed 100 percent of the Mexican bean beetles and 15 percent of the aphids at concentrations of '1000 and 250 parts per million, respectively.

. EXAMPLE 26 The perhalocoumalin derivative of the formula CH: ==O

exhibited complete control of Pythium and Rhizoctonia at a concentration of 64 pounds per acre. At concentrations of 255 and 38 parts per million, respectively, it exhibited complete control of the bacteria Staphylococcus aureus (Gram and Xanthomonas phascoli (Guam EXAMPLE 27 The perhalocoumalin derivative of the formula exhibited complete control of Rhizoctonia, Staphylococcus aureus (Gram Pseudomonas phaseolicola (Gram and Xanthomonas phascoli (Gram at concentrations of 64 pounds per acre, 255 parts per million, 76 parts per million, and 76 parts per million, respectively.

EXAMPLE 28 The perhalocoumalin derivative of the formula.

exhibited complete control of Sclerotium, Staphylococcus aureus, (Gram Pseudomonas phaseolicola (Gram and Xanthomonas phascoli (Gram at concentrations of 64 pounds per acre, 255 parts per million, 76 parts per million, and 255 parts per million, respectively. It killed 100 percent of the mites at a concentration of 250 parts per million, and it killed 50 percent of the houseflies within 24 hours at a concentration of 1000 parts per million.

EXAMPLE 29 The perhalocoumalin derivative of the formula SCH-4H2 exhibited complete control of Pythium and Sclerotium at a concentration of 64 pounds per acre.

EXAMPLE 30 The perhalocoumalin derivative of the formula exhibited complete control of Sclerotium at a concentra 12 tion of 64 pounds per acre. It killed percent of the houseflies Within two hours and 100 percent of the mites at a concentration of 1000 parts per million.

EXAMPLE 31 The perhalocoumalin derivative of the formula exhibited complete control of Pythium, Rhizoctonia, Sclerotium, and Staphylococcus aureus (Gram at concentrations of 64 pounds per acre, 64 pounds per acre, 64 pounds per acre, and 255 parts per million, respectively,

EXAMPLE 32 The perhalocoumalin derivative of the formula ll S-P-OCHa OCH:

exhibited complete control of Staphylococcus aureus (Gram and Pseudomonas phaseolicola (Gram at concentration of 255 and 76 parts per million respectively. It killed 42 percent of the mites at a concentration of 1000 parts per million.

EXAMPLE 33 The perhalocoumalin of the formula The perhalocoumalin derivative of the formula exhibited complete control of Fusarium and Pseudomonas phaseolicola (Gram at concentrations of 64 pounds per acre and 255 parts per million, respectively. It killed 50 percent of the aphids at a concentration of 1000 parts per million.

While there have been described various embodiments of the present invention, the methods and compositions described are not intended to be understood as limiting the scope of the invention as it is realized that changes therein are possible. It is further intended that each element recited in any of the following claims is to be understood as referring to all equivalent elements for accomplishing substantially the same results in substantially the wherein X is a halogen selected from the group consisting of chlorine and bromine;

(R) Z is a substituent in which the following relationship exists:

When n is- Z is- Bromine, iodine, fluorine. 1 Oxygen, sulfur. 2 Nitrogen. 3 Carbon.

and wherein R is selected from the group consisting of hydrogen; straight or branched chain alkyl of 1 to 6 carbon atoms provided that Z is sulfur; (c) cyclohexyl; (d)

phenyl or benzyl; (e) CH CH OR or --CH CH SH when Z is oxygen or sulfur.

2. The method of claim 1 wherein R is oxygen.

3. The method of claim 1 wherein Z is nitrogen.

4. The method of claim 1 wherein Z is oxygen and R is an alkyl group of from 6 to 18 carbon atoms.

5. The method of claim 1 wherein n is 0, Z is iodine, and X is chlorine.

6. The method of claim 1 wherein R is phenyl or benzyl.

7. The method of claim 1 wherein R is C H CH OH or -CH CH SH.

8. The method of claim 1 wherein Z is sulfur.

9. The method of claim 8, wherein a bactericidal amount of said compound is applied to the locus of bacteria.

10. The method of claim 8, wherein a fungicidal amount of said compound is applied to the locus of fungi.

11. The method of claim 8, wherein a miticidal amount of said compound is applied to the locus of mites.

12. The method of claim 8 wherein X is a halogen.

13. The method of claim 8 wherein n is 1.

14. The method of claim 8 wherein R is hydrogen.

15. The method of claim 8 wherein X is chlorine.

16. The method of claim 8 wherein R is an alkyl group of 1 to 18 carbon atoms.

References Cited UNITED STATES PATENTS 3,122,557 2/1964 Molho 424279 SAM ROSEN, Primary Examiner UNITED STATES PATENT OFFICE CERTIFICATE CF CORRECTION Patent No. 3,755,59l Dated August 2 973 Inventor(s) Victor Mark and Leon Zenqierski It is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:

olumn l, line 27, Abstract "a substi tuents" should read ---a substi tuent---;. line 66, that portion of the formula reading OH H 0 II I i ll H -C C should read -C C I l Z(R) v Z(R) Column 3, line 37, "centrigrade" should read ---centi grade---. Column lines l, 2 and 3,, delete "using one nucleophi l e precursor, and then subject the mono-substi tu ted perhalocoumal in"; line l5, that portion of the formula reading ."base HCl should read -base HCl+--. Column 5, Table I, under Base, Example 2, (C H N" should read ---(C H N--; Table 1, under Base, Example 3, .".((;2H2) N" should read ---(C2H5)3N- Table I, under Base, Example 4-, "(C2H2)3N" should read ---(C2H5)3N---; Table I, under Base, Example 5, "(C H NH" should read .---(C2H5)2NH--; Table I, under Base, Example 6, ."'(C2H2)3N" should read (C2H5)3N--; Table I, under Base, xample 7, C H N" should read ---(C H N-.--;- Table I, under Base, Example 8, "(C2H2)2NH should read ---(C H NH--; I Table I, under Base Example 9, "(C H N" should read ---'(C H l N Table I, under Cl, Example 2, "3,20%3 2 should read "-37.0535. 'Table I, under Empirical Formula, xample "C17.L H 7 Cl O 7S" should read ---C .L,H 5,7Cl 0 8-"; Table I, under Formula, Example 5, "(C2H5) 7NH" should read --(C H5)2NH---; Table I, Example 8, under Empirical Formula, "C H Cl N 0 should read ---C17H 1Cl N302---; Table I, under H, Example 9, "l ,7/+.7"- should read ---l.7/l .7---;- Table I, under 8, Example l l "lOJT/l 0.2" should read --'-l0.0/lO.'2---. Column 6, l ine 60, the formula should appear as fol lows 0SCH3 L I 1 .J

UNITED STATES PATENT OFFICE (5/69) Page 2 CERTIFICATE OF CORRECTION Patent No. 3,155, 59l Dated August 28, 1973 Inventor) Victor Mark and Leon Zengierski it is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:

Column 7, line 17, "taken" should read ---take---; line 69, "back" should read ---black---. Column 9, line 53, "and the" should read ---and at the---; line 67, after "aureus" insert ---(Gram Pseudomonas phaseol icol a (Gram and---. Column 10, l ine +5, "The derived coumalin product" should read ---The perhalocoumalin derivative---. Column 12,

line 32, "concencr'ation' should read --concentrations--; line 60, the formula should appear as fol lows OC6H5 C Column 13, line 30, before 'hydrogen" insert ---(a)---, after semicolon insert ---(b)-. Column IL}, line 1, after "-CH CH SH" insert a line 2, delete.

Signed and sealed this 7 th day of May 197M.-

(SEAL) Attest:

EDWARD I'LFLETCHZ ERJR. C. MARSHALL DANN Attesting Officer I Commissioner of Patents 

